An engineered membrane-bound guanylyl cyclase with light-switchable activity

BMC Biology

Georg Nagel¹  Shiqiang Gao¹  Yuehui Tian² 

[1] Department of Neurophysiology, Institute of Physiology, Biocenter, University of Wuerzburg, 97070, Wuerzburg, Germany;Department of Neurophysiology, Institute of Physiology, Biocenter, University of Wuerzburg, 97070, Wuerzburg, Germany;Present address: Environmental Microbiomics Research Center, School of Environmental Science and Engineering, Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Sun Yat-sen University, 510006, Guangzhou, China;

关键词: Chlamydomonas reinhardtii;    Guanylyl cyclase;    Optogenetics;    Rhodopsin;    Cyclic GMP;   

DOI  :  10.1186/s12915-021-00978-6

来源: Springer

【 摘 要 】

BackgroundMicrobial rhodopsins vary in their chemical properties, from light sensitive ion transport to different enzymatic activities. Recently, a novel family of two-component Cyclase (rhod)opsins (2c-Cyclop) from the green algae Chlamydomonas reinhardtii and Volvox carteri was characterized, revealing a light-inhibited guanylyl cyclase (GC) activity. More genes similar to 2c-Cyclop exist in algal genomes, but their molecular and physiological functions remained uncharacterized.ResultsChlamyopsin-5 (Cop5) from C. reinhardtii is related to Cr2c-Cyclop1 (Cop6) and can be expressed in Xenopus laevis oocytes, but shows no GC activity. Here, we exchanged parts of Cop5 with the corresponding ones of Cr2c-Cyclop1. When exchanging the opsin part of Cr2c-Cyclop1 with that of Cop5, we obtained a bi-stable guanylyl cyclase (switch-Cyclop1) whose activity can be switched by short light flashes. The GC activity of switch-Cyclop1 is increased for hours by a short 380 nm illumination and switched off (20-fold decreased) by blue or green light. switch-Cyclop1 is very light-sensitive and can half-maximally be activated by ~ 150 photons/nm2 of 380 nm (~ 73 J/m2) or inhibited by ~ 40 photons/nm2 of 473 nm (~ 18 J/m2).ConclusionsThis engineered guanylyl cyclase is the first light-switchable enzyme for cGMP level regulation. Light-regulated cGMP production with high light-sensitivity is a promising technique for the non-invasive investigation of the effects of cGMP signaling in many different tissues.

【 授权许可】

CC BY